Wakle Monali, Khole Vrinda
Gamete Immunobiology Department, National Institute for Research in Reproductive Health, J.M. Street, Parel, Mumbai, India.
Biochem Biophys Res Commun. 2007 Mar 9;354(2):528-34. doi: 10.1016/j.bbrc.2007.01.021. Epub 2007 Jan 12.
Using post-vasectomy monoclonal antibody we recently identified a testis specific sperm auto-antigen called TSA70 which is post-meiotically expressed and plays a role in sperm motility and capacitation-acrosome reaction. In the present study, we report its cytoskeletal nature based on its resistance to various high ionic salt solutions. TSA70 is developmentally regulated and appears postpubertally. The two protein spots identified by 2D WB namely TSA1-pI=5.821, MW=77.050 and TSA3-pI=6.173, MW=75.519 showed sequence homology to Cenexin/odf2 indicating that two are isoforms of the same protein. The immunoreactivity of TSA70 with anti-Cenexin antibody substantiates its homology with Cenexin/odf2. In silico analysis revealed the presence of two leucine zippers in TSA70 and also predicted potential phosphorylation sites at serine, threonine, and tyrosine residues. The phosphorylated status of TSA70 was further confirmed by immunoblot analysis. The differential cellular expression suggests that TSA70 is a novel member of Cenexin/odf2 family that exhibits functional divergence.
利用输精管切除术后单克隆抗体，我们最近鉴定出一种睾丸特异性精子自身抗原，称为TSA70，它在减数分裂后表达，在精子运动和获能-顶体反应中发挥作用。在本研究中，我们基于其对各种高离子盐溶液的抗性报告了其细胞骨架性质。TSA70受发育调控，在青春期后出现。二维蛋白质免疫印迹法鉴定出的两个蛋白点，即TSA1-pI = 5.821，MW = 77.050和TSA3-pI = 6.173，MW = 75.519，显示出与Cenexin/odf2的序列同源性，表明两者是同一蛋白的同工型。TSA70与抗Cenexin抗体的免疫反应证实了它与Cenexin/odf2的同源性。计算机分析显示TSA70中存在两个亮氨酸拉链，还预测了丝氨酸、苏氨酸和酪氨酸残基处的潜在磷酸化位点。免疫印迹分析进一步证实了TSA70的磷酸化状态。不同的细胞表达表明TSA70是Cenexin/odf2家族的一个新成员，具有功能差异。
